Process for producing an extract of proteolytic enzyms.



TED STATES PA Bron.

'IOMENOSUKE MU'I'O, OF KCBE, JAPAN, ASSIGNOR '10 KANEGAFUCHI BOSEKIIKABUSHIKI KWAISHA, 'OF TOKYO-EU, JAPAN.

PROCESS FOR PRODUCING AN EXTRACT OF PROTEOLYTIC ENZYMS.

No Drawing.

a specification.

My invention relates to the process of ext'racting proteolytic enzymfrom the bodies of silkworms and several other cocoon-forming insects atany desired stage of their growth, as well as from their secretions anddischarges, and has for its object to produce a proteolytic enzym easilyand economically,

v and thereby to produce the same in amounts other cocoon-formin usefulin industrial and medicinal uses.

Silkworms and several other cocoon-forming insects, in theirmetamorphoses from pupa into moth, each secrete a certain juice by whicha portion of the cocoon is softened and loosened. Considering that thissoftening and loosening is due, not to a physical action of wetting, butto a chemical action of certain ingredients contained in the secretion,the inventor. has devoted himself K to the investigation of saidsecretion, and has now discovered therein a certain proteolytic enzymwhich is capable of softening sericin and thereby loosening fibers ofsilk. Furthermore, the inventor has invented a process of extractingproteolytic enzym not only from the secreted juice but also from thebodies of silkworms and other cocoon-forming insects at every stage oftheir growth, and from their castofi skins,'secretions and discharges,all of which materials were heretofore considered as mere waste insericulture and filature.

carrying the invention into effect, to any products of silkworms,containing the enzym in (giestion, e. g. silkworm pupae, silkworm-mot s,their cast-01f skins, secretlons and discharges, or similar productsfrom chestnut worms, pine worms, or various insects (hereinafterreferred to under t e eneral ression products of cocoon-forming insectsthere is preferably added glycerin or other'antiseptic materialincapable of in urlng the enzym, and the same then added to distilledwater at a tem erature of 5 to 15 C. and allowed to stan for six toforty-eight hours,

or the material is mixed with, and allowed Specification of LettersPatent.

an Extract of Pro-' teol'ytic Enzyms, of which the following is"Patented Mar. 4:, 1919.

Application filed April 25, 1917. Serial No. $4,343.

to stand in distilled water at a temperature of. 40 C. for five tofifteen minutes, and

then the. material squeezed. The extract, thus obtalned, is put in avacuum evaporator pan, and it is condensed to 10% to 20% of its volume,while at a temperature of about C., and the sediment is then re. moved.

In the extraction and concentration of the enzym, it will be obviousthat temperatures should be employed which do not injuriously affect theenzym. The temperatures above stated are safe'in this respect.Temperatures above' 0. should be avoided for reasons stated below.

According to the purpose for which the product is to be used, the saidproduct may be then, further condensed in vacuum to a paste or a solid.Moreover, a comparatively pure residue containing the enzym may beproduced by the application thereto of an agent capable of precipitatingthe enzym, such as alcohol, ether, sulfate of ammonia, etc., or bysalting-out.

The residue produced by one of the above processes, contains a largequantity of a certain proteolytic enzym which decomposes several kindsof albumin, especially sericin. Sericin becomes readily dissolved in ashort timeby the action of this proteolytic enzym.'

The other valuableproperties of this proteolyticenzym are :that it ismost active at a-temperature of 35 to 40 C.;'that it is resistant todryv heat, in that it can be subjected, in a 'dry state, to atemperature of 100 0., without entirely losing its activity;

while in moist heat,'its activity is gradually Now, as this extract ofproteolytic enzym may dissolve sericin in a short time, it ma be usedeffectively in filature, waste si spmmng, and manufacture of floss-silk,to

soften or dissolve sericin and to loosen fibers of silk, and if refined,the extract'ma b used advantageously for the finishing o silk threadsand silk fabrics. Furthermore, this extract may be applied, like otherproteo lytic enzyms, in medicine, as well as in the temperature of notmaterially above C preparation of other purposes.

It is awell known fact that various proteolytic enzyms hitherto knownmay be used for the above mentioned purposes, but most of thoseheretofore known are too expensive to be generally used for industrialpurposes.

According to the present invention, howmixture with water at atemperature not materially above 40 0., separating the liquor from theundissolved matter, and

then separating the enzym from the liquor.

2. A process of producing a proteolytic enzym which comprises digestingmaterial containing products of cocoon-forming insects with water at atemperature not materially above 40 (1., separating the liquor from theundissolved matter, and then separating the enzym from the liquor.

3. A process ofproducing a proteolytic enzym from products ofcocoon-forming in-. sects which comprises adding an antiseptic,

material not capable of materially injuring the enzym to said material,digesting the mixture with waterat a temperature not materially above4010, separating the liquor from the undissolved matter, concentratingthe liquor in vacuum to about 10 to 20% of its original volume, while ata separating the enzym from the and then liquor.

enzym from products of cocoon-forming peptone, in tanning, and for 4. Aprocess of producing ,a proteolytic v memes insects which com risesadding an antiseptic material not capa le of materially mjuring theenzym to said material, digesting the mixture with water at atemperature not materially above 40 0., separating the liquor from theundissolved matter, concentrating the liquor in vacuum to about 10 to20% of its original volume, while at a temperature of not materiallyabove 45 C., and then precipitating the enzym in a substantially pure.state, from the concentrated extract.

5. A process of producing a proteolytic enzym from cocoon-forming insectmaterial which comprises digesting such material with glycerin and waterat a temperature too low to materially injure the proteolytic enzym,separating the liquor from the undissolved matter, and separating theenzym from the liquor.

6. A process of producing aproteolytic enzym which comprises digestingmaterial containing products of cocoon-forming insects with an aqueousliquid at a temperature too low to injure the proteolytic enzymcontained therein, separating the liquor from the undissolved matter,and separating the enzym from the liquor.

7. The herein described new proteolytic enzym which may be produced fromproducts of cocoon-forming insects, which enzym is capable of dissolvingsericin, and other proteid bodies, iscapable in a dry state of beingheated to 100 (3., without loslng its activity, is incapable of beingheated in a wet state to 80 0. without losing a large part of itsactivity, and which enzym is active in weak acid solutions and inneutral solutions.

In testimony whereof I have 'afiixed my signature in presence of twowitnesses.

- TOMENOSUKE MUTO.

Witnesses:

EISHIRO ABE, Y. NAGAMATIS'U.

